Phospho p44 42 mapk erk1 2 rabbit monoclonal antibody

Cytoplasmic cell extracts were separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes for western blotting. Membranes were incubated with p44/42 MAPK (ERK1/2) rabbit monoclonal antibody (Cell Signaling Technology, Danvers, USA), Phospho-p44/42 MAPK (ERK1/2) rabbit monoclonal antibody (Cell Signaling Technology), or anti β-actin antibody (Sigma-Aldrich, St. Louis, USA) at 4°C overnight, as appropriate. After washing with Tris-buffered saline (TBS) containing 0.02% (v/v) Tween 20, membranes were incubated with horseradish peroxidase-conjugated secondary antibody (Sigma-Aldrich) at 37°C for 1 h. Immunoreactions were then visualized using the enhanced chemiluminescence detection system (UVP Inc., Upland, USA).

PBK/TOPK mouse monoclonal antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), phospho-p44/42 MAPK (Erk1/2) rabbit monoclonal antibody, phospho-AKT (Ser473) rabbit monoclonal antibody, phospho-PTEN (ser380/Thr382/382) mouse monoclonal antibody, and EpCAM (VU1D9) mouse monoclonal antibody conjugated to Alexa Fluor 594 were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Cytokeratin 19 rabbit monoclonal antibody was purchased from Abcam, Inc. (Cambridge, MA, USA), CD45 rabbit monoclonal antibody was purchased from Biosen, Inc. (Beijing, China), goat anti-mouse IgG conjugated to FITC and TRITC were purchased from Biosen, Inc. (Beijing, China), goat anti-rabbit IgG conjugated to FITC and TRITC were purchased from Biosen Inc. (Beijing, China), HRP-labeled goat anti-mouse IgG(H+L) and goat anti-rabbit IgG(H+L) were purchased from SinoBio, Inc. (SuZhou, China), and 4,6-diamino-2-phenyl indole (DAPI) was purchased from Beyotime, Inc. (Beijing, China), Diaminobenzidine (DAB) substrate was purchased from Dako Denmark A/S (Glostrup, Denmark), packaging vectors including pMD2.0G and psPAX were purchased from Addgene Inc. (Cambridge, MA, USA), polybrene were purchased from EMD Millipore (Billerica, MA, USA), puromycin were purchased from Sigma-Aldrich (St Louis, MO, USA).